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当前位置: 首页 > 文章 > 奶牛乳腺上皮细胞的体外培养及蛋白含量测定 畜牧与兽医 2014,46 (9) 11-14
Position: Home > Articles > In vitro culture of bovine mammary epithelial cell and determination of its protein content Animal Husbandry & Veterinary Medicine 2014,46 (9) 11-14

奶牛乳腺上皮细胞的体外培养及蛋白含量测定

作  者：
罗莹;吴金节;王希春;马良友;刘红梅;张胜军
单  位：
安徽农业大学动物科技学院
关键词：
奶牛;乳腺上皮细胞;β-酪蛋白;细胞培养
摘  要：
为建立稳定的奶牛乳腺上皮细胞系(BMEC),进一步研究乳腺上皮细胞体外蛋白的分泌情况,本试验首先采用组织块贴壁法获得奶牛乳腺上皮细胞,用胰蛋白酶纯化细胞,并运用SDS-PAGE进行鉴定。采用MTT分析细胞活力,瑞氏-吉姆萨染色法对细胞核进行分析,并测定细胞上清液中β-酪蛋白(β-CN)含量。结果显示:本试验成功培养出较纯化的奶牛乳腺上皮细胞;细胞生长曲线呈明显"S"型,用瑞氏-吉姆萨法染色细胞,细胞核形态多为卵圆形且呈现明显蓝紫色。原代乳腺上皮细胞可连续多次传代且未出现永生化,细胞呈单层贴壁生长,证明细胞活力良好;测定其β-CN含量为3.3 mg/L。
译  名：
In vitro culture of bovine mammary epithelial cell and determination of its protein content
作  者：
LUO Ying;WU Jinjie;WANG Xichun;MA Liangyou;LIU Hongmei;ZHANG Shengjun;College of Animal Science and Technology,Anhui Agricultural University;
关键词：
cow;;epithelial mammary cells;;β-casein;;cell culture
摘  要：
The aim of this study is to establish a stable bovine mammary epithelial cell( BMEC) line and to determine the protein secretion of BMEC. The primary culture of BMEC was performed using tissue explant method and purified by trypsin digestion method. Cell identification was carried out by SDS-PAGE analysis,cell viability was determined by MTT,and the nuclei were observed by Wright-Giemsa staining. The level of β-casein was determined by capillary electrophoresis. The result indicated that the purified BMEC was successfully obtained. The cell growth curve showed a typical"S"shape,and wright-Giemsa staining showed that the nuclei were oval and obvious bluepurple. The primary mammary epithelial cells could be passaged several times continuously and immortalized cells did not appear. The mammary epithelial cells grew in monolayer. The content of β-casein was 3. 3 mg /L.

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